Development and Application of a gp60-Based Subtyping Tool for Cryptosporidium bovis

Cryptosporidium bovis is a common enteric pathogen in bovine animals. The research on transmission characteristics of the pathogen is hampered by the lack of subtyping tools. In this study, we retrieve the nucleotide sequence of the 60 kDa glycoprotein (GP60) from the whole genome sequences of C. bovis we obtained previously and analyze its sequence characteristics. Despite a typical structure of the GP60 protein, the GP60 of C. bovis had only 19.3–45.3% sequence identity to those of other Cryptosporidium species. On the basis of the gene sequence, a subtype typing tool was developed for C. bovis and used in the analysis of 486 C. bovis samples from dairy cattle, yaks, beef cattle, and water buffalos from China. Sixty-eight sequence types were identified from 260 subtyped samples, forming six subtype families, namely XXVIa to XXVIf. The mosaic sequence patterns among subtype families and the 121 potential recombination events identified among the sequences both suggest the occurrence of genetic recombination at the locus. No obvious host adaptation and geographic differences in the distribution of subtype families were observed. Most farms with more extensive sampling had more than one subtype family, and the dominant subtype families on a farm appeared to differ between pre- and post-weaned calves, indicating the likely occurrence of multiple episodes of C. bovis infections. There was an association between XXVId infection and occurrence of moderate diarrhea in dairy cattle. The subtyping tool developed and the data generated in the study might improve our knowledge of the genetic diversity and transmission of C. bovis.

[1]  Lihua Xiao,et al.  Subtyping Cryptosporidium xiaoi, a Common Pathogen in Sheep and Goats , 2021, Pathogens.

[2]  Lihua Xiao,et al.  Development of a Subtyping Tool for Zoonotic Pathogen Cryptosporidium canis , 2020, Journal of Clinical Microbiology.

[3]  Kuankuan Zhang,et al.  Longitudinal detection of Cryptosporidium spp. in 1–10-week-old dairy calves on a farm in Xinjiang, China , 2020, Parasitology Research.

[4]  N. Huang,et al.  Subtyping Cryptosporidium ryanae: A Common Pathogen in Bovine Animals , 2020, Microorganisms.

[5]  Lihua Xiao,et al.  Comparative genomic analysis of three intestinal species reveals reductions in secreted pathogenesis determinants in bovine-specific and non-pathogenic Cryptosporidium species , 2020, Microbial genomics.

[6]  Longxian Zhang,et al.  Population structure and geographical segregation of Cryptosporidium parvum IId subtypes in cattle in China , 2020, Parasites & Vectors.

[7]  M. Santín Cryptosporidium and Giardia in Ruminants. , 2020, The Veterinary clinics of North America. Food animal practice.

[8]  R. Chalmers,et al.  Development of a gp60-subtyping method for Cryptosporidium felis , 2020, Parasites & Vectors.

[9]  Lihua Xiao,et al.  Outbreak of cryptosporidiosis due to Cryptosporidium parvum subtype IIdA19G1 in neonatal calves on a dairy farm in China , 2019, International Journal for Parasitology.

[10]  U. Emanuelson,et al.  Infection dynamics of Cryptosporidium bovis and Cryptosporidium ryanae in a Swedish dairy herd , 2019, Veterinary parasitology: X.

[11]  Lihua Xiao,et al.  Prevalence and genotypic identification of Cryptosporidium spp., Giardia duodenalis and Enterocytozoon bieneusi in pre-weaned dairy calves in Guangdong, China , 2019, Parasites & Vectors.

[12]  Lihua Xiao,et al.  Genetic Diversity and Population Structure of Cryptosporidium. , 2018, Trends in parasitology.

[13]  T. Orro,et al.  Cryptosporidium outbreak in calves on a large dairy farm: Effect of treatment and the association with the inflammatory response and short-term weight gain , 2017, Research in Veterinary Science.

[14]  M. Grigg,et al.  Genomics and molecular epidemiology of Cryptosporidium species. , 2017, Acta tropica.

[15]  Wenchao Yan,et al.  Subtype analysis of zoonotic pathogen Cryptosporidium skunk genotype. , 2017, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[16]  Lihua Xiao,et al.  Molecular Epidemiology of Cryptosporidiosis in China , 2017, Front. Microbiol..

[17]  B. S. Sandhu,et al.  Periurban outbreaks of bovine calf scours in Northern India caused by Cryptosporidium in association with other enteropathogens , 2017, Epidemiology and Infection.

[18]  E. Houpt,et al.  Cryptosporidium and Giardia Infections in Children: A Review. , 2017, Pediatric clinics of North America.

[19]  Lihua Xiao,et al.  Longitudinal monitoring of Cryptosporidium species in pre-weaned dairy calves on five farms in Shanghai, China. , 2017, Veterinary parasitology.

[20]  Jong Wan Kim,et al.  Multilocus typing of Cryptosporidium spp. in young calves with diarrhea in Korea , 2016, Veterinary Parasitology.

[21]  Wenxian Wu,et al.  Distribution of Cryptosporidium species in Tibetan sheep and yaks in Qinghai, China. , 2016, Veterinary parasitology.

[22]  C. R. Stensvold,et al.  Development and Application of a gp60-Based Typing Assay for Cryptosporidium viatorum , 2015, Journal of Clinical Microbiology.

[23]  Chunfu Yang,et al.  Subtyping Novel Zoonotic Pathogen Cryptosporidium Chipmunk Genotype I , 2015, Journal of Clinical Microbiology.

[24]  Wenxian Wu,et al.  Occurrence and molecular characterization of Cryptosporidium spp. and Enterocytozoon bieneusi in dairy cattle, beef cattle and water buffaloes in China. , 2015, Veterinary parasitology.

[25]  R. Fayer,et al.  Cryptosporidium species in humans and animals: current understanding and research needs , 2014, Parasitology.

[26]  R. Fayer,et al.  Subtyping Cryptosporidium ubiquitum,a Zoonotic Pathogen Emerging in Humans , 2014, Emerging infectious diseases.

[27]  Lihua Xiao,et al.  Multilocus Sequence Typing of an Emerging Cryptosporidium hominis Subtype in the United States , 2013, Journal of Clinical Microbiology.

[28]  Lihua Xiao,et al.  Population genetic characterisation of dominant Cryptosporidium parvum subtype IIaA15G2R1. , 2013, International journal for parasitology.

[29]  C. Björkman,et al.  Is there a need for improved Cryptosporidium diagnostics in Swedish calves? , 2012, International Journal for Parasitology.

[30]  Takako Sasaki,et al.  Molecular characterization of Cryptosporidium spp. in grazing beef cattle in Japan. , 2012, Veterinary parasitology.

[31]  U. Zahid,et al.  Drug combination therapy in control of cryptosporidiosis in Ludhiana district of Punjab , 2012, Journal of Parasitic Diseases.

[32]  U. Ryan,et al.  Molecular characterization of Cryptosporidium and Giardia in pre-weaned calves in Western Australia and New South Wales. , 2011, Veterinary parasitology.

[33]  D. Pethick,et al.  Cryptosporidium andersoni in Western Australian feedlot cattle. , 2010, Australian veterinary journal.

[34]  J. Mattsson,et al.  Molecular characterisation of Cryptosporidium isolates from Swedish dairy cattle in relation to age, diarrhoea and region. , 2010, Veterinary parasitology.

[35]  R. Fayer,et al.  A longitudinal study of cryptosporidiosis in dairy cattle from birth to 2 years of age. , 2008, Veterinary parasitology.

[36]  S. Tzipori,et al.  Molecular Cloning and Expression of a Gene EncodingCryptosporidium parvum Glycoproteins gp40 and gp15 , 2000, Infection and Immunity.

[37]  J. Gut,et al.  Cloning and Sequence Analysis of a Highly Polymorphic Cryptosporidium parvum Gene Encoding a 60-Kilodalton Glycoprotein and Characterization of Its 15- and 45-Kilodalton Zoite Surface Antigen Products , 2000, Infection and Immunity.

[38]  R. Fayer,et al.  Genetic Diversity within Cryptosporidium parvum and Related Cryptosporidium Species , 1999, Applied and Environmental Microbiology.

[39]  E. Esteban,et al.  Cryptosporidium muris: prevalence, persistency, and detrimental effect on milk production in a drylot dairy. , 1995, Journal of dairy science.